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Combination Metabolomics Approach for Identifying Endogenous Substrates of Carnitine/Organic Cation Transporter OCTN1.
Pharm Res. 2018 Oct 02; 35(11):224.PR

Abstract

PURPOSE

Solute carrier SLC22A4 encodes the carnitine/organic cation transporter OCTN1 and is associated with inflammatory bowel disease, although little is known about how this gene is linked to pathogenesis. The aim of the present study was to identify endogenous substrates that are associated with gastrointestinal inflammation.

METHODS

HEK293/OCTN1 and mock cells were incubated with colon extracts isolated from dextran sodium sulfate-induced colitis mice; the subsequent cell lysates were mixed with the amino group selective reagent 3-aminopyridyl-N-hydroxysuccinimidyl carbamate (APDS), to selectively label OCTN1 substrates. Precursor ion scanning against the fragment ion of APDS was then used to identify candidate OCTN1 substrates.

RESULTS

Over 10,000 peaks were detected by precursor ion scanning; m/z 342 had a higher signal in HEK293/OCTN1 compared to mock cells. This peak was detected as a divalent ion that contained four APDS-derived fragments and was identified as spermine. Spermine concentration in peripheral blood mononuclear cells from octn1 gene knockout mice (octn1-/-) was significantly lower than in wild-type mice. Lipopolysaccharide-induced gene expression of inflammatory cytokines in peritoneal macrophages from octn1-/- mice was lower than in wild-type mice.

CONCLUSIONS

The combination metabolomics approach can provide a novel tool to identify endogenous substrates of OCTN1.

Authors+Show Affiliations

Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan.Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan.Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan.Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan.Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan.Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan.Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki, Japan.Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan.Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan. ykato@p.kanazawa-u.ac.jp.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30280275

Citation

Masuo, Yusuke, et al. "Combination Metabolomics Approach for Identifying Endogenous Substrates of Carnitine/Organic Cation Transporter OCTN1." Pharmaceutical Research, vol. 35, no. 11, 2018, p. 224.
Masuo Y, Ohba Y, Yamada K, et al. Combination Metabolomics Approach for Identifying Endogenous Substrates of Carnitine/Organic Cation Transporter OCTN1. Pharm Res. 2018;35(11):224.
Masuo, Y., Ohba, Y., Yamada, K., Al-Shammari, A. H., Seba, N., Nakamichi, N., Ogihara, T., Kunishima, M., & Kato, Y. (2018). Combination Metabolomics Approach for Identifying Endogenous Substrates of Carnitine/Organic Cation Transporter OCTN1. Pharmaceutical Research, 35(11), 224. https://doi.org/10.1007/s11095-018-2507-1
Masuo Y, et al. Combination Metabolomics Approach for Identifying Endogenous Substrates of Carnitine/Organic Cation Transporter OCTN1. Pharm Res. 2018 Oct 2;35(11):224. PubMed PMID: 30280275.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Combination Metabolomics Approach for Identifying Endogenous Substrates of Carnitine/Organic Cation Transporter OCTN1. AU - Masuo,Yusuke, AU - Ohba,Yuri, AU - Yamada,Kohei, AU - Al-Shammari,Aya Hasan, AU - Seba,Natsumi, AU - Nakamichi,Noritaka, AU - Ogihara,Takuo, AU - Kunishima,Munetaka, AU - Kato,Yukio, Y1 - 2018/10/02/ PY - 2018/04/02/received PY - 2018/09/18/accepted PY - 2018/10/4/entrez PY - 2018/10/4/pubmed PY - 2019/7/3/medline KW - Crohn’s disease KW - OCTN1 KW - endogenous substrates KW - metabolomics KW - transporter SP - 224 EP - 224 JF - Pharmaceutical research JO - Pharm Res VL - 35 IS - 11 N2 - PURPOSE: Solute carrier SLC22A4 encodes the carnitine/organic cation transporter OCTN1 and is associated with inflammatory bowel disease, although little is known about how this gene is linked to pathogenesis. The aim of the present study was to identify endogenous substrates that are associated with gastrointestinal inflammation. METHODS: HEK293/OCTN1 and mock cells were incubated with colon extracts isolated from dextran sodium sulfate-induced colitis mice; the subsequent cell lysates were mixed with the amino group selective reagent 3-aminopyridyl-N-hydroxysuccinimidyl carbamate (APDS), to selectively label OCTN1 substrates. Precursor ion scanning against the fragment ion of APDS was then used to identify candidate OCTN1 substrates. RESULTS: Over 10,000 peaks were detected by precursor ion scanning; m/z 342 had a higher signal in HEK293/OCTN1 compared to mock cells. This peak was detected as a divalent ion that contained four APDS-derived fragments and was identified as spermine. Spermine concentration in peripheral blood mononuclear cells from octn1 gene knockout mice (octn1-/-) was significantly lower than in wild-type mice. Lipopolysaccharide-induced gene expression of inflammatory cytokines in peritoneal macrophages from octn1-/- mice was lower than in wild-type mice. CONCLUSIONS: The combination metabolomics approach can provide a novel tool to identify endogenous substrates of OCTN1. SN - 1573-904X UR - https://www.unboundmedicine.com/medline/citation/30280275/Combination_Metabolomics_Approach_for_Identifying_Endogenous_Substrates_of_Carnitine/Organic_Cation_Transporter_OCTN1_ L2 - https://doi.org/10.1007/s11095-018-2507-1 DB - PRIME DP - Unbound Medicine ER -